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1 Institut de Recherches Servier, 92150 Suresnes, France; and 2 Cardiovascular Research Center and Department of Medicine, Massachusetts General Hospital and Harvard Medical School, Charlestown, Massachusetts 02129
Nitric oxide (NO) is a potent vasodilator, but
it can also modulate contractile responses of the airway smooth muscle.
Whether or not endothelial (e) NO synthase (NOS) contributes to the
regulation of bronchial tone is unknown at present. Experiments were
designed to investigate the isoforms of NOS that are expressed in
murine airways and to determine whether or not the endogenous release of NO modulates bronchial tone in wild-type mice and in mice with targeted deletion of eNOS [eNOS(
/)]. The presence of neuronal NOS
(nNOS), inducible NOS (iNOS), and eNOS in murine trachea and lung
parenchyma was assessed by RT-PCR, immunoblotting, and
immunohistochemistry. Airway resistance was measured in
conscious unrestrained mice by means of a whole body
plethysmography chamber. The three isoforms of NOS were
constitutively present in lungs of wild-type mice, whereas only iNOS
and nNOS were present in eNOS(/) mice. Labeling of nNOS was
localized in submucosal airway nerves but was not consistently
detected, and iNOS immunoreactivity was observed in tracheal and
bronchiolar epithelial cells, whereas eNOS was expressed in endothelial
cells. In wild-type mice, treatment with N-nitro-L-arginine methyl ester, but not with
aminoguanidine, potentiated the increase in airway resistance produced
by inhalation of methacholine. eNOS(/) mice were hyperresponsive to
inhaled methacholine and markedly less sensitive to
N-nitro-L-arginine methyl ester. These results
demonstrate that the three NOS isoforms are expressed constitutively in
murine lung and that NO derived from eNOS plays a physiological role in
controlling bronchial airway reactivity.
epithelium; immunohistochemistry; methacholine; endothelial nitric oxide synthase knockout mice; N-nitro-L-arginine methyl ester; airway reactivity; reverse transcription-polymerase chain reaction; Western blot
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